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1、ArchVirol(2006)151:20852094DOI10.1007/s00705-006-0769-0AssemblyofHCVE1andE2glycoproteinsintocoronavirusVLPsBriefReportN.Jourdan1,2,G.-J.Godeke3,M.Penaud2,G.Mottola1,A.Sorrentino1,P.J.M.Rottier3,andS.Bonatti11DipartimentodiBiochimicaeBiotecnologieMediche,Universit`adiNapoliFe
2、dericoII,Napoli,Italy2LaboratoiredeTh´erapieG´enique,INSERMUMRU649,CHUHotelDieu,Nantes,France3VirologyDivision,DepartmentofInfectiousDiseasesandImmunology,VeterinaryFaculty,Utrecht,TheNetherlandsReceivedJanuary3,2006;acceptedMarch9,2006PublishedonlineMay3,2006cSpringer-Verl
3、ag2006Summary.HepatitisCvirus(HCV)isbelievedtoassemblebybuddingintomembranesoftheearlysecretorypathway,consistentwiththemembranelocationwheretheviralenvelopeglycoproteinsE1andE2accumulatewhenexpressed.Coronavirusassemblyalsotakesplaceatpre-Golgimembranes.Here,wegener-atedcor
4、onavirus-likeparticlescarryingintheirenvelopechimericHCVglycopro-teinscomposedoftheectodomainsofE1andE2,eachfusedtothetransmembraneplusendodomainofthemousehepatitiscoronavirusspikeglycoprotein.ThechimericparticlesystemwillenablestructuralandfunctionalstudiesoftheHCVglycoprot
5、eins.∗HepatitisCvirus(HCV)isthemajoretiologicalagentofhumannon-A,non-Bhepatitis[4,17]andhasinfectedmillionsofpeopleworldwide[20].Thisflaviviruscauseschronicliverdiseaseforwhichtreatmentoptionsarelimitedandnovaccineisyetavailable.Thestudyofitsbiologyhasbeenseverelyhamperedbyth
6、elackofviralculturesystems,asituationthathasrecentlychangedbythegenerationofinfectiousHCVgenomesthatreplicateefficientlyincellculture[21,37,38].LittleisstillknownaboutthebiogenesisandstructureofHCVparticles.Thevirusiscomposedofacore(C)proteinthatpackagesthepositive-strandedRN
7、Agenomeandenvelopeproteins,E1andE2,whicharegeneratedbycleavage2086N.Jourdanetal.fromthesingleprecursorpolyprotein,translatedfromthegenomicRNA.E1andparticularlyE2areheavilyN-glycosylatedproteinsanchoredinmembranesbytheirC-terminalhydrophobicdomain.Expressionstudieshaveshownth
8、eglycoproteinstoberetainedinearlycompartmentsofthesecretorypathway,consiste
